ISSN: 2320-5407 Int. J. Adv. Res.

9(06), 681-690

Journal Homepage: - epage: - www.journalijar.com

Article DOI: 10.21474/IJAR01/13063

DOI URL: http://dx.doi.org/10.21474/IJAR01/13063

RESEARCH ARTICLE
APPLICATIONS OF MOLECULAR MARKERS AND GENOMIC TOOLS IN PEARL MILLET
[CENCHRUS AMERICANUS (L.) MORRONESYN. PENNISETUM GLAUCUM (L.) R. BR.]

Bassirou Mbacké and Abdoulaye Dieng

École Nationale Supérieure d’Agriculture/UniversitéIba Der Thiam de Thiès BP 296 Thiès/Sénégal.
……………………………………………………………………………………………………....
Manuscript Info Abstract
……………………. ………………………………………………………………
Manuscript History
Received: 23 April 2021
Final Accepted: 25 May 2021
Published: June 2021
Key words:-
Pearl Millet, QTL Mapping, Stress
Tolerance, Genetic Diversity, GBS
Thisreviewsummarizesthemainmolecularmarkersandtheirapplicationso
n pearl milletaswellasasummaryof the
discoveriesonitsreferencegenome.Molecularmarkers,unlikemorphologic
alandbiochemicalmarkers,arehighlypolymorphicandneutral.
Theirgreatliabilitycomesfromthefactthattheydirectlyconcern the
DNA.Theyhavebeenwidelyusedonpearl millet,rangingfromlow
andmedium-throughputtohigh-throughput markers,
targetingspecificregionsorcharacterizinggermplasmat thegenomelevel.
Many
studiesrelatetomappingusingdifferentpopulationsandhaveidentifiedQTL
slinkedtoimportantagronomictraits(floweringtime,tolerancetodrought,to
mildew,phosphorus absorption),iron
content...Studieshavealsobeenconductedondomesticationsyndromeands
howedtheir importance of genes
flowfromwildmilletstocultivatedvarieties. Genotyping-by-Sequencing -
a rapid, cost-effective and reduced representation sequencing method –
has been used to assess genetic diversity, population structure, LD and
heterotic pool formation in pearl millet. A draft genome sequence that
can serve as a reference for further development of genomics-assisted
breeding is now available. It is an important milestone in generating
genomic resources for pearl millet. Annotation of 24,000 genes
indicates that enrichment of wax biosynthesis genes providing potential
genetic mechanisms for heat and drought tolerance.
Althoughmolecularmarkersarewidelyappliedtomillet,geneticandgenomi
cresourcesarestilllimitedcomparedtootherimportantcereals.However,the
availabilityofacollectionofinbredlinesrepresentativeofgermplasmandare
ferencegenomeoffernewperspectivesintheimprovement of pearl millet.

Copy Right, IJAR, 2021,. All rights reserved.

……………………………………………………………………………………………………....
Introduction:-
Pearl milletis the staple food crop for millions of people in semi-arid areas of Africa and Asia. Its grain has an
important nutritional value. Its fodder has good digestibility and stubble is used in construction (fence, huts). Faced
with the threat of food insecurity following climate change, pearl millet appears to be a cereal to be promoted, due to
its high tolerance to drought [1]. It can produce in hot areas where rainfall is very low and tolerates soils with high
pH and salinity [2]. The strong allogamy resulting from a pronounced protogyny is the source of a high level of
heterozygosity. This genetic variability is an interest but also a challenge for obtaining homogeneous and

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CorrespondingAuthor:- Bassirou Mbacké
Address:- École Nationale Supérieure d’Agriculture/Université de Thiès BP 296 Thiès/Sénégal.
ISSN: 2320-5407 Int. J. Adv. Res. 9(06), 681-690

reproducible research material. Various research institutes have developed molecular markers that have made it
possible to have important genetic resources and to establish a reference genome for millet.This article presents a
review on the main genetic markers, their applications and the results obtained on this species, followed by a
summary presentation of the pearl millet genome.

Main molecular markers

Molecular markers are very polymorphic, neutral, reliable and in almost unlimited numbers. They are independent
of the environment and the stage of development of the plant. A molecular marker is a polymorphic locus whose
genotype provides information on the genotype of neighboring loci. Allelic variability at the marker locus should
ideally not have effects other than those that determine its genotype. Depending on the detection method and the
throughput, the molecular markers can be divided into three groups: low-throughput markers, based on restriction
fragment length polymorphism (RFLP), medium-throughput markers based on PCR: RAPD, AFLP, SSR..., high-
throughput markers based on amplicon polymorphism/restriction fragments (DArT) or nucleotide sequence
polymorphism (SNP). Apart from SSRs, low- and medium-throughput markers have now been little used since the
advent of high-throughput sequencing. We review them here to take into account that high-throughput next-
generation sequencing (NGS) tools are not available everywhere.

RFLP
If the DNA of two individuals differs, its digestion by different restriction enzymes will generate fragments of
different lengths. Single-stranded DNA fragments are separated by electrophoresis, transferred and fixed to a
membrane. A restriction polymorphism is shown by hybridization of complementary molecular probes, followed by
an X-ray; the probe-enzyme pair is a marker. RFLP are co-dominant and reproducible markers. However, their
detection is long and laborious; the technique is almost no longer used.

RAPD
It makes possible to detect polymorphism in the absence of information on amplified sequences. The amplification
is carried out from a random sequence primer that hybridizes with the sequences that are complementary to it. If two
sites of hybridization are close and in the opposite direction, PCR allows amplification. If the DNA of two
individuals differs at the hybridization sites, a polymorphism of presence / absence of bands can be demonstrated on
an electrophoresis gel. Although these dominant markers can detect polymorphic loci, they are neither locus-specific
nor reproducible.

AFLP
It is based on the joint revelation of restriction polymorphisms and hybridization of arbitrary primers. The first step
is to cleave the DNA by two different enzymes and bind the fragments to complementary adapters. PCR
amplification is performed using primers of sequences homologous to those of adapters extended in 3 '. The primers
are oriented so that only fragments having a different restriction site at each end are amplified. The bands are
visualized by acrylamide gel electrophoresis. AFLP are dominant and non-locus specific. Although they are
anonymous, their level of reproducibility and sensitivity are high. However, the method is laborious and is not
automatable.

SSR
Simple Sequence Repeats or microsatellites are tandem repeated sequences of nucleotide patterns distributed
throughout the genome. Their polymorphism is based on the variation in the number of repetition units.
Microsatellites are very polymorphic, codominant and locus-specific; the technique is automatable and does not
require a large amount of DNA. These advantages mean that, despite a high detection cost, these markers are still
widely used.

DArT
Diversity Array Technology is based on selective amplification of a subset of amplicons obtained by digesting
genomic DNA with a pair of restriction enzymes. The fragments are labeled and hybridized to a DNA chip. A
collection of previously identified polymorphic amplicons are fixed on the chip. If an individual produces a
particular amplicon, it will recognize its complement on the chip and it will result in a positive signal. If the
amplicon is absent, no hybridization signal is obtained at this position. The technique is reproducible and does not
require a large amount of DNA but the markers are dominant.

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SNP
Single Nucleotide Polymorphisms have proven to be the markers of choice in genetics and plant breeding with the
emergence of high-throughput sequencing technologies and platforms, accompanied by significant cost reductions.

Definition and characteristics in plants

An SNP represents a polymorphism of nucleotides between alleles at a locus. SNPs are the most important source of
variability between individuals at the molecular level. Polymorphism can be a mutation by transversion or transition
or an insertion/deletion (InDel). In the case of changes in two nucleotides or InDel-type events of some nucleotides,
one speaks of "polymorphisms of simple nucleotides" [3]. Most of the variability in quantitative traits is related to
SNPs. Theoretically polymorphism can concern the four nucleotide variants. In practice, SNPs are generally bi-
allelic and variations take place at different frequencies [4]. The weakness of SNP polymorphism due to bi-allelism
can be compensated by their higher frequency [5]. In coding sequences, SNPs can be synonymous and not alter the
amino acid sequence. If they are not synonymous, significant polymorphisms resulting from differences in amino
acid composition can be identified. The frequency of an SNP is given according to the frequency of the minor
alleles; an SNP with a minor allele frequency C of 0.40 implies that 40% of a population has the C allele compared
to the more common allele which is found in 60% of the population.

Applications
The increasing importance given to SNPs comes from their applications in population mapping and characterization.
It is assumed that most of the quantitative traits are due to SNPs not yet identified. Their abundance and the rapid
evolution of genotyping technologies make it possible to generate new genetic maps or saturate existing ones. As
mentioned earlier, the limited information associated with their bi-allelic nature is compensated by a high frequency;
thus, a map of 700 to 900 SNPs was equivalent to a map of 300 to 400 SSRs [6].

Genotyping technologies
Genotyping of SNP involves discriminating alleles at a given locus based on the change in one of the four
nucleotides. A large number of techniques and platforms are available [7]. Most technologies are based on
hybridization, enzymatic methods or the physical properties of DNA. Those based on DNA hybridization probes
complementary to SNP sites include: dynamic hybridization of specific allele, molecular tags and biochips. The
principle of a biochip is the convergence of DNA capture on a solid surface, its hybridization and visualization
under a microscope. There are two main approaches: first, a single primer extension hybrid upstream of the SNP, a
DNA polymerase incorporates a complementary labeled ddNTP; secondly, a specific allele primer extension
corresponding to the variant is extended by PCR [7]. The melting temperature and conformation of a single strand of
DNA are the basis for allele discrimination in technologies based on the physical properties of DNA. These include
the single-strand conformation polymorphism and the temperature gradient on electrophoresis gel. There are many
other technologies and platforms including genotyping-by-sequencing (GBS) which uses restriction enzymes and
has the advantage of incorporating the simultaneous discovery of SNP and genotyping of individuals.

Main applications in pearl millet

Important molecular genetic resources have been obtained on millet. The main applications are reviewed here.

Genetic map
Several genetic maps have been established using mainly self-fertilized populations or fixed recombinant lines [8].
The first map drawn up with RFLP consisted of markers spread over seven linkage groups (LG) [9]. The analysis of
nineteen genotypes with 200 DNA probes from the F2 progeny highlighted the very high genetic variability of
millet. This map had a density of 181 loci for a coverage of 303 cM. A second map improved the first one with the
combined use of 353 RFLP markers and 65 SSR markers [10]. The number of mapped loci increased to 242 with a
coverage of 473 cM. This was followed by a more saturated genetic map from fixed recombinant lines, consisting of
258 DArT markers and 63 SSR markers, with 321 loci [11]. Although this map was larger (1148 cM), the number of
markers obtained by population was less. The discovery of microsatellites in expressed regions has made it possible
to develop EST-SSR markers in a simple and straightforward way from EST databases. This combination allows the
selection of markers according to the physiological or biochemical properties expressed. A consensus map was
obtained using a combination of 99 EST-SSR markers on F7 populations of recombinant lines [12]. This study
improved coverage and filled in the gaps on previous maps.

The emergence of new high-throughput sequencing technologies represents an important step in genetic mapping.
This is how GBS was developed and proven its effectiveness in important crops such as corn and barley [13], [14].
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In millet, GBS involving a combination of PstI-MspI restriction enzymes resulted in 3,321 SNP markers that were
used to establish a more saturated map, with a population of 93 F2 individuals from a cross between the SOSAT-
IBL197 parental lines and PS202-14 [8]; 314 SNP markers distributed evenly over seven linkage groups (LG) made
it possible to build a map covering a genetic distance of 640 cM, with an average interval of 2.1 cM between
markers. To bridge the gap between this map and the previous ones, 19 SSR markers are analyzed on the parents of
the population; four of them were polymorphic and made it possible to establish a correspondence of four LG with
the map of Qi et al. [10]. It is also with the GBS that the most saturated genetic map was obtained on millet [15].
99% of it, developed using 150 recombinant lines, has intervals between neighboring markers of less than 5 cM.
This map was compared with that of Rajaram et al. [12]: 191 markers were correctly aligned and 16 concordant but
on different sites. The existence of a reference genome for millet offers the possibility of aligning the 4900 SNPs
identified. Comparing the lengths of LG with the map obtained by Moumouni et al. [8], the distances between
markers were more extensive on some LG. The differences observed between a previously established map [15] can
be explained by the use of the Kosambi distance, unlike other maps based on the Haldane distance. The adaptability
of millet to various constraints makes it a model species for tolerance studies. A collection called PMiGAP (Pearl
Millet inbred Germplasm Association Panel) comprising 346 lines was created from about 1000 cultivars,
accessions and relatives of mapping population from Africa and Asia. This panel offers new perspectives in the
mapping of QTL. Linkage mapping using experimental populations is to track the transmission of QTL alleles and
markers over generations. The segregation of markers in the population is related to the genotypic variability
according of the phenotypes observed. A phenotypic difference suggests the segregation of alleles into a QTL bound
to this marker. Advances in high-throughput SNP sequencing make association mapping a valuable tool increasingly
used in plants [16]. It is an alternative or complementarity to linkage mapping. It can be divided into two categories:
gene-candidate association mapping and genome-wide association studies (GWAS) that focus on the analysis of
several phenotypes in statistically significant relation to SNPs over the entire genome. Here we present some
examples of mapping applications of QTLs that are of great agronomic importance in millet.

Flowering time
The duration from sowing to flowering is an important factor in the genotype-environment interaction for yield [17].
The precocity of this trait allows millet to complete its cycle in conditions of water stress. A phenotype-genotype
association methodology has been developed with the aim of identifying genes involved in the variation of this trait
using SSR and AFLP markers; this study made it possible to associate the PHYC and MADS11 genes with the
duration of flowering [18]. These associations have been validated by QTL analyses that suggest that
polymorphisms within the PHYC gene may explain the variation in flowering time. Another study on the MADS11
block also identified a PgMADS11 polymorphism associated with the variation of this trait [19]. A mapping of
agronomic traits including flowering time in a population of recombinant lines allowed the detection of six
flowering-related QTLs on five chromosomes; a major QTL on LG3 was common to the duration of flowering and
the height of the plant [20]. In addition, a map showed a QTL related to flowering and resistance to pyriculariosis
[15].

Downy mildew tolerance

Caused by Sclerosporagraminicola, this disease causes significant damage to millet. Yield losses can be as high as
80%, with a decrease in the quality of grains and fodder [21]. Tolerance to downy mildew is quantitatively heritable
[22]. QTL of high-effect resistance were detected on LG1, LG6 and LG7 against strains from India, on LG4 for
strains from Niger and Nigeria and on LG2, LG6 and LG7 for pathogens from Senegal [23]. Some of these QTLs
were systematically identified in repeated screenings but none were effective on all strains. Nine putative QTLs, one
of which was common to the eight pathogen populations from Africa and Asia, were detected [24]. Mapping based
on SSR markers using an F8 population of recombinant lines demonstrated five QTLs with a broad effect on
tolerance to three isolates of the pathogen from India, at the seedling stage and in a controlled environment [21]. The
introgression assisted by resistance marker saves time compared to the conventional method. An association of ISSR
and SCAR markers was used to assist in the improvement of the parental lines of the HHB 67 hybrid for resistance
to Downy mildew [17]. This combination of markers was also analyzed on a collection of millet; BAND 863 SCAR-
ISSR was identified on all seven resistant lines and was absent on all five susceptible lines [25]. Although linkage
mapping identified a number of important QTLs in millet, this method was limited by the resolution provided.
Association mapping based on linkage disequilibrium (LD), considers more alleles and gives a greater resolution.
Thus, a GWAS was conducted with 77 genotypes to identify SNP markers related to resistance to downy mildew but
the results have to be validated [26].

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Rust and pyriculariosis tolerance

Mapping using DArTs and SSRs on an F7 population of recombinant lines was performed to identify QTLs related
to rust resistance caused by Puccinia substriata var. indica [11]. The map was used to identify a QTL on LG1
believed to confer stable rust resistance in India. A map has been validated for its usefulness in mapping QTLs
related to resistance to pyriculariosis (Pyricularia grisea (Cke.) Sacc.) [15].

Drought tolerance
Drought tolerance is a complex trait strongly influenced by the environment and controlled by many genes [27].
Physio-morphological traits include changes in root and vegetative characteristics for efficient water absorption and
conservation in the plant. Among the most significant factors, we can note: conductance, stay green, density and root
depth, roughness and cuticular thickness, osmotic adjustment, accumulation of stress proteins... Pearl millet has a
tillering potential that allows it to compensate for an early water deficit. However, it is difficult to accurately assess
drought tolerance. Several studies have focused on the harvest index (HI). High HI has been shown to be related to
the ability of tolerant varieties to conserve water during the vegetative phase and use it for grain filling; a QTL
associated with phenotype was identified and introduced by backcrossing [28]. QTL related to delayed senescence,
an SNP in a gene encoding acetyl CoA carboxylase linked to yield and harvest index, as well as an InDel in a gene
bound to a chlorophyll synthesis protein significantly associated with stay green and water stress yield were mapped
[29]. A QTL related to reduced transpiration in the vegetative stage was co-mapped with another of terminal drought
tolerance [29]. Under salinity conditions, drought tolerance has been observed to be linked to a reduction in salt
absorption and accumulation in leaves [30]. A study that aimed to identify the genetic components involved in early
drought tolerance was conducted on 188 West African lines phenotyped under early water stress and non-limiting
irrigation [31]. A GWAS carried out with 392,493 SNPs generated by GBS led to the identification of QTL
controlling biomass production under conditions of early water stress and stay green. Genes involved in the
synthesis of sirohaem and waxes co-localize respectively with these two QTLs [31].

Phosphorus absorption
In West Africa, pearl millet is grown on soils that are often very low in phosphorus. DArTs have identified nine
markers significantly associated with phosphorus-related traits [32]. Some polymorphisms were divided between
traits related to phosphorus absorption efficiency and yield. However, validation of these markers is necessary to
determine their applicability in improvement programs.

Iron and zinc contents

On a population of recombinant lines from a cross where one parent has high iron and zinc levels, DArT and SSR
markers were used to establish QTLs related to the content of these elements in millet [20]. The analysis revealed 11
QTLs related to the iron content in the grains and eight related to zinc. Three high-impact QTLs for these elements
were co-mapped in the population. Favorable alleles are transmitted by the male parent with the highest levels of
iron and zinc. Growing pearl millet varieties enriched with these elements will be valuable in areas where rural
populations face malnutrition.

Genetic diversity
Accurate assessment of diversity is of great importance in the analysis of cultivar genetic variability, introduction of
genes of interest, phylogeny or population structuring. Having evolved under environmental constraints, millet has a
high level of polymorphism, both between and within accessions. Before the emergence of molecular markers,
methods based on pedigree data, morphological, biochemical characteristics...have been used in diversity studies in
millet [33]. Markers such as RFLP [35], AFLP [36], RAPD [37], SSR [38], [39], [40] and a combination of
DArT/SSR [11] have been used in several diversity studies. Previously used polymorphic SSRs were used for
neutral characterization, followed by adaptive genotyping characterization of the PgMADS11 and PgPHYC genes
involved in flowering to assess the genetic diversity and adaptation of a collection of millet from Senegal [41]. The
analysis shows a great diversity and differentiation into two pools between early flowering cultivars and those late
flowering, according with the zones of culture. The adaptive diversity assessment shows the specificity of a
PgPHYC gene SNP and an InDel at the PgMADS11 gene associated with the observed differences in flowering
durations.

Genome-wide characterization of germplasm is necessary for a better understanding of millet genomic resources.
Also, the genetic diversity of 248 cultivars from Senegal and 252 accessions from a global collection has been
analyzed by GBS with a couple of PstI-MspI restriction enzymes that allowed the identification of 83,875 SNPs
[42]. Senegalese cultivars had the highest levels of genetic diversity while accessions from Southern Africa and Asia
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had the lowest levels of diversity (Figure 1). A clear structuring of the population between the varieties of Senegal
and the world collection on the one hand, and between countries within the world collection on the other hand, was
observed. A weak population structure was noted within cultivars from Senegal (Figure 2). In addition, the decrease
in the linkage disequilibrium (LD) was much rapid and delayed.The rapid decreaseof the LD and the lack of
structuring along the agro-ecological zones of Senegalese millet open up possibilities for association mapping. A
comparison with the genomes of Setariaitalica and Sorghum bicolor indicates large regions of synteny and large-
scale rearrangements in millet evolution.

Fig 1:Genome-wide estimates of genetic diversity. AComparison of nucleotide diversity among accessions from different countries. The “**” sign
represents a significant difference (P < 0.01) between Senegalese landraces and the accessions from the given countries. BThe distribution of
inbreeding coefficients for global accessions and Senegalese landraces (Hu etal., 2015)

Fig2:Population structure.Different colors represent the different subpopulations from ADMIXTURE, and text under the
barplot indicates the origins of the corresponding germplasm. The K value left of the figures indicates the assumed number of
subpopulations (Hu etal., 2015)

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Another application of the GBS generated 82,112 SNPs that were also used to analyze genetic diversity, population
structuring and LD on a collection of millet, following the alignment of reads on the reference genome [43]. The
study reveals the structuring into six groups (with genetic differentiation within them) and a more rapid decrease in
LD in the West African subpopulation

Other applications
The phenomenon of heterosis has been studied a lot on many species. In millet, SSR markers were used to identify
heterotic groups in a collection of hybrid parental lines, with the reference genotype Tift23 D2B1-P1-P5. as control
[44]. These markers have made it possible to distinguish two groups that should be refined by increasing the number
of crosses and to evaluate the hybrids obtained in more representative locations.The origin and the areas of
domestication of millet have been the subject of much investigation. Whole-genome sequencing (WGS) data from
221 accessions consisting of wild forms and traditional varieties representative of the geographical diversity of
millet and the modelling of allele flows have confirmed the West African origin of the species and the syndrome of
its domestication south of the Sahara, about 190 years ago 4900 years [45]. The study shows that millet varieties
grown in the world derive from common ancestors originating in the Sahelian Center. During its diffusion, the
cultivated varieties would have hybridized with local wild millet, leading to the great genetic diversity observed in
the Western and Eastern areas of the Sahel. The hypothesis of gene flow from wild populations during
domestication in these regions was confirmed by the identification of 15 genomic regions that were the subject of
adaptive introgression. Previous studies had hypothesized that Senegal and eastern Sahel could constitute centres of
domestication of millet [42]; [46]. Double-use millet (grain and fodder) is of increasing interest to research. SSRs
and GBS have been used on hybrid varieties to identify markers related to forage quality [47]. Between two
successive cuts, a positive evolution in crude protein content and in-vitro digestibility of organic matter was
observed.

Pearl millet genome

Thegenomesizeofpearl millethasbeenestimatedat4.0pg/2C[48].Theentiregenomeofthereferencegenotype
Tift23D2B1-P1-
P5wassequenced[2].1.49Tbofsequencingdatawereassembledinto1.58Gbofcontigs,or90%ofthegenome.TheaverageG
Ccontent(47.9%)ishigherthanthatofsorghum,barleyorrice.1.22Gbofrepeatedsequenceshavebeenidentified,oratleast77.
2%ofthegenome.Combiningtranscriptomic
dataandbioinformaticsanalyses,itispredictedthatthemilletgenomecontainssome38,579genesofwhich96.7%havebeenan
notatedandfunctionsattributedto72.30%ofthese.Comparisonofgenefamilieswithothercerealgenomesindicatesenrichm
entinsequencesinvolvedinthebiosynthesisofcutin,suberinandwax;compoundsconstitutingbarrierstowaterlossbytheplan
t[2];[49]. Thus,theexpansionofthesegenefamilieswouldcontributetomillet'stolerancetodrought.
TheidentificationofaQTLoftolerancetoearlywaterstressassociatedwithageneinvolvedinthebiosynthesisofthesecompou
ndshelpstoreinforcethisidea[31].
Themajorityofpathogenresistancegenesinplantscontainanucleotidebindingsite(NBS).
TheidentificationofgenescontainingNBSmakesitpossibletoidentifyputativeresistancegenes.Ofthe378NBSgenespresen
tinthemilletgenome,360couldbemappedonatleastoneofthesevenchromosomes.994linesandaccessionsincluding345lin
esfromthePMiGAPandincluding31wildaccessionswerere-
sequencedinordertobetterunderstandthepopulationstructure,geneticdiversityanddomesticationofmilletbytheWGRS,G
BSandSSRapproaches,with450,000SNPsidentified.Fourmaingroupsstandout:threebringtogetherthewild(correspondin
gtotheEast,CentralandWestAfricanregions)andone,thecultivatedvarieties.Theclosestwildaccessionstocultivarscomefr
omWestAfrica;thisreinforcestheWestAfricanoriginofmillet.Thestrongpopulationstructureobservedinwildaccessionsre
flectsageneticdiversitystillinsufficientlyexploited[43].Theweakstructuringofcultivatedvarietiessuggestsahomogeneou
sdiversitylinkedtotheunhinderedselectionandspreadofmilletinlargecultivatedregionsofAfricaandAsia.Followingseque
ncingandre-sequencingwork,88,256primersfor74,891
microsatelliteswereidentifiedonthegenome,aswellas29,542,173SNPsonthePMiGAPcollection;1054trait-
markersfor15agronomictraitshavebeenidentified,aswellasasetofgenefamilies that
allowabetterunderstandingofmillet'sexceptionaltolerancetoheatanddrought.

Conclusion:-
The objective of this reviewwas to synthesize the resources obtained with molecular markers on pearl millet. This is
how we presented the main genetic linkage maps established. Pearl millet evolved in Africa and Asia under difficult
environmental conditions that gave it an adaptation to these environments. However, it still faces many biotic and
abiotic constraints that limit its production of grain and quality fodder. Many QTLs related to interesting agronomic
traits have been mapped. Genomic tools have been used to know the syndrome of domestication of millet in the
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south of the Sahara and have shown the importance of genes introgressionfrom wild populations to cultivated
varieties. Molecular markers have also provided a more accurate assessment of millet genetic diversity and
phylogeny relationships. The strong population structure observed in wild accessions reflects a genetic diversity that
deserves to be further exploited. The availability of a reference genome presents an important resource and offers
immense prospects for improving this important cereal.

Acknowledgements:-
We thank Laurent LAPLAZE3 for correcting the manuscript.
3. UMR/DIADE, Institut de Recherches pour le Développement, 911 Av. AGROPOLIS 34395 Montpellier CEDEX
5, France.

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